Expression, purification, and circular dichroism analysis of human CDK9

Author(s):Show less -

Leopoldino, Andreia Machado ; Canduri, Fernanda ; Cabral, Hamilton ; Junqueira, Magno ; Marqui, Alessandra Bernadete Trovó de ; Apponi, Luciano H. ; Fonseca, Isabel Osório da ; Domont, Gilberto Barbosa ; Santos, Diógenes S. ; Valentini, Sandro ; et al.

• Document type: Journal article
• Source: Protein Expression and Purification; v. 47, n. 2, p. 614-620, June 2006.
• Field of knowledge: Biological Sciences - Genetics

Abstract

The human cyclin-dependent kinase 9 (CDK9) protein was expressed in E. coli BL21 using the pET23a vector at 30 °C. Several milligrams of protein were purified from soluble fraction using ionic exchange and ATP-affinity chromatography. The structural quality of recombinant CDK9 and the estimation of its secondary structure were obtained by circular dichroism. Structural models of CDK9 presented 26% of helices in agreement with the spectra by circular dichroism analysis. This is the first report on human CDK9 expression in Escherichia coli and structure analysis and provides the first step for the development of CDK9 inhibitors. (AU)

• FAPESP's process: 01/07532-0 - Structural genomics of cyclin dependent kinases and plant defensive proteinases and their natural inhibitors
• Grantee: Walter Filgueira de Azevedo Junior
• Support Opportunities: Regular Research Grants